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The role of growth factors for neovascularization of CNS tumor

生长因子在中枢神经系统肿瘤新生血管形成中的作用

基本信息

批准号：
10670177
负责人：
SAWADA Tatsuo
金额：
$ 1.41万
依托单位：
Tokyo Women's Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

项目摘要

1) Expression of VEGH, VEGFR and VEGFmRNA in human and rat brain tymumor.Vascular endothelial growth factor (VEGF) is a secreted angiogeneic regulator or growth factor of blood vessels during embryonic development and in tumors. We performed expression of VEGF, VEGFR - 1 and VEGFmRNA in human and rat brain.To evaluate to the role of VEGF in the development of vacular system in tumor, and the effects of the microcircumstance, we analyzed existence VEGF, VEGFR(flt-1) and expression of VEGF mRNA using immunohistochemistry and in situ hybridization in human brain tumors and rat trasnsplanted tumors. We used surgically removed astrocytic tumors and metastatic tumors, subcutaneous and intracerebral transplanted animals with ENU induced rat glioma cultured cell (C6). In astrocytic tumors and metastatic tumors, expression of VEGF and VEGER were recognized in newly-formed micro blood vessels which form glomeruloid proliferations. But no expression in endothelial cells without glomeruloid prolif … More erations. In transplanted tumors, no significant differences of proliferative activities of tumor cells and the morphlogy of newly-formed micro blood vessels were identified in each tumors. No glomeruolid proliferations were noted in newly formed micro blood vessels in both tumors.Expression of VEGF and VEGFR were recognized in the endothelial cells but no expression of VEGF mRNA. In cerebral tumor, tumor infiltrations were noted in perivascular infilatrarions was recognized in subcutaneous tumor, pseduopalisading patterns were recognized in the subcutaneous ones and the expression of VEGF mRNA were increased in the tumor cells of pseudopalisading areas. The transplanted tumors were though to be affected by of transplanted areas. The biological and morpholgical characteristics the transplanted tumors were thought to be affected by the microcircumstance of transplanted areas.2) Expression in VEGF, VEGFR, and VEGFmRNA in rat vasculognesis.To evaluate to the role of VEGF in the development of vascular system in normal physiological state, we were analyzed the existence VEGF and expression of VEGF mRNA using immunohistochemistry and in situ hybridization in newborn rats. 24 newborn rat brains were surgically removed at 1, 3, 5, 7, 10 and 14 days after the birth. The expression of VEGF mRNA was recognized in the endothelial cells of capillaries from one day to the 7 days after the birth, the maximum expressions were demonstarated at 10 days. The expression is decreased at 10 days and no expressions are recognized at 14 days after birth. VEGFmRNA was up regulated in the neurons of all specmens, especially in Purukinje cells and neuron at hippocampi. Maximal expressions are recognized at 5days after birth and decreased grandually. Immunohistochemical stainngs using anti VEGF monoclonal antibody obtained similar findings these findings highly suggested that autocrine and paracrine signal transduction for the stimulation of the growth of neovasculature were established in rat vasculogenesis.3) The relationship of tight junction related protein and tumor proliferative actoivities.To elucidate the neovasculature of brain tumors preserve the blood brain barrier (BBB) functions, we studies the expressions of the MDR1 gene products, P-glycoprotein(PgP) and tight juinction related protein. Zo-1 using immunohistochemistry. 30 brain tumors were examined using an anti PgP and an anti ZO-1 Mab, and comparing the expression of VEGF, VEGFR and PCNA.. Positive reactions for PGP and Zo-1 were recoginzed in the endothelial cells in newly formed micro-blloc vessels in primariy glioma but not those in the meningiomas. Although endothelial cells in newly formed micro-blood vessels of all metastatic carcinomas showed positive reaction; negative reactions were seen in those of the primary carcinomas. Weak reactions for anti PgP and Zo-1 compared to those of endothelial cells forming glomeruloid proliferations in newly formed micro-blood vessles in glioblastomas and at the border of the surrounding cerebral tissue of metastatic carcinomas. The negative relations between positive reactions for BBB related proteins, and the expression of VEGF and the proliferative activities were recognized. These findings suggested that the endothelial cells in newly formed micro-blood vessels preserved the fundtions of BBB. Less

1)VEGH、VEGFR和VEGFmRNA在人和大鼠脑肿瘤中的表达。血管内皮生长因子(VEGFR)是胚胎发育和肿瘤中分泌的血管生成调节因子或生长因子。为探讨血管内皮生长因子在肿瘤血管系统发育中的作用及微环境的影响，我们采用免疫组织化学和原位杂交的方法，分析了血管内皮生长因子、血管内皮生长因子受体(VEGFR，Flt-1)在人脑肿瘤和大鼠移植瘤中的表达。我们使用手术切除的星形细胞瘤和转移性肿瘤，皮下和脑内移植的动物与ENU诱导的大鼠胶质瘤培养细胞(C6)。在星形细胞肿瘤和转移性肿瘤中，VEGF和Veger在新生微血管中表达，形成肾小球增殖。但在无肾小球样长…的血管内皮细胞中无表达更多的颂词。在移植瘤中，肿瘤细胞的增殖活性和新生微血管的形态均无明显差异。两种肿瘤的新生微血管均未见肾小球样增生，血管内皮细胞表达血管内皮生长因子和血管内皮生长因子受体，但未见血管内皮生长因子基因的表达。在脑肿瘤中，肿瘤在皮下肿瘤可见血管周围浸润，皮下肿瘤可见假扁平排列，假扁平区肿瘤细胞中血管内皮生长因子基因表达增强。移植瘤受移植区域大小的影响。移植瘤的生物学和形态特征受移植区微环境的影响。2)新生大鼠血管新生血管中血管内皮生长因子及其受体的表达。为探讨血管内皮生长因子在正常生理状态下血管系统发育中的作用，采用免疫组织化学和原位杂交方法检测新生大鼠血管内皮细胞生长因子的存在和血管内皮生长因子基因的表达。24只新生大鼠分别于出生后1、3、5、7、10、14天手术切除脑组织。生后1d至7d，毛细血管内皮细胞可检测到VEGFmRNA的表达，10d时表达最强。出生后10天表达减弱，14天后无表达。VEGFmRNA在所有物种的神经元中表达上调，尤其是Purukinje细胞和海马神经元。出生后5天表达最强，以后逐渐下降。用抗血管内皮生长因子的单抗进行免疫组织化学染色，得到了类似的结果。这些结果高度提示在大鼠的血管生成中建立了自分泌和旁分泌的信号转导机制。3)紧密连接相关蛋白与肿瘤增殖活性的关系。为了阐明脑肿瘤新生血管保护血脑屏障(BBB)的功能，我们研究了mdr1基因产物、P-糖蛋白(Pgp)和紧密连接相关蛋白的表达。ZO-1免疫组织化学染色。应用抗Pgp单抗和抗ZO-1单抗对30例脑肿瘤进行检测，并比较其在肿瘤组织中的表达。Pgp和Zo-1在原发胶质瘤新生微血管内皮细胞中呈阳性反应，而在脑膜瘤中未见阳性反应。所有转移性癌新生微血管内皮细胞均呈阳性反应，而原发癌则呈阴性反应。与内皮细胞相比，抗Pgp和Zo-1的反应较弱，在胶质母细胞瘤的新生微血管和转移性癌周围脑组织中形成肾小球样增生。BBB相关蛋白阳性反应与血管内皮生长因子表达及细胞增殖活性呈负相关。提示新生微血管内皮细胞对血脑屏障功能具有保护作用。较少